Disintegrin Molecules to Target Glioblastoma Integrins with High Affinity Design, Characterization and Integration into Silica-Based Nanocarrier Systems
Project no. 21792 · Status: Kapanmış (Yayın bekleniyor) · 2020
Summary
<p>Within the scope of the presented project, amino acid sequences of disintegrin structures containing bioactive RGD sequence isolated from the venom of our country's endemic V. anatolica were investigated by our group. In molecular docking and molecular dynamics studies with cyclic pentapeptide structures derived from these sequences, the binding of alpha and beta 3 disintegrin of sequences containing two aspartic acids such as RGDDN and GRGDD and sequences containing an unnatural amino acid γ-carboxyglutamic acid (Gla). showed that their interactions with Mn 2+ ions from the RGD region may be better than the RGD sequence.</p><p><br></p><p> In the light of this information, new cyclic pentapeptide sequences containing the above mentioned sequences will be designed. The interactions of these newly designed cyclic peptides with integrins will be investigated by molecular modeling studies. Five of the disintegrins, which were observed to have the best scores and interactions as a result of molecular modeling studies, will be selected and synthesized by solid phase peptide synthesis method and then purified. The structures of the purified disintegrin molecules will be determined by NMR and LC-MS/MS and taken to the bioactivity screening test (cell proliferation-cytotoxicity test). Studies will be conducted in comparison with ecistatin disintegrin. For this purpose, higher amounts of cyclic peptide/peptides with the best activity will be synthesized according to the data to be obtained, and they will be taken into further biological characterization tests (cell adhesion test, determination of integrin specificities by flow cytometry and confirmation by western blot, determination of anti-angiogenesis effect). In order to pave the way for therapeutic applications of the polypeptides, whose biological characterization tests were carried out at the last stage of the study, they will be loaded with fluorescent dye-doped mesoporous silica nanoparticles (MSN) and with the help of nanocarrier systems, it will be ensured that the bio-active substances show their effectiveness even in low amounts by overcoming the blood-brain barrier, which prevents drug delivery to the brain. . The project is unique in that it is the first study in this field in our country.</p>